Multilocus sequence typing MLST with high solution sensitivity and specificity is widely used to study the population genetic structure of pathogen by amplification and sequencing of the housekeeping genes. MLST also provides more evidence and plays an important role in parasite research. This paper reviews the principle and method of MLST and its applica-tion on population genetic structure analysis of parasites.

Objective To investigate the clinical efficacy of Cerus and cucumis polypeptide injection in the prevention of heterotopic ossification for the drug treatment of traumatic knee joint fracture after operation.Methods To retrospect the analysis about 150 cases with fracture reduction and internal fixation of traumatic around the knee joint fracture from January 2010 to January 2014 in Daliang Hospital of Shunde District of Foshan,including 89 cases of reduction and internal fixation were performed in group A,and with safflower injection infusion therapy,61 cases with allergic constitution were not given by Cerus and cucumis polypeptide injection in group B.According to the activity degree and HSS scoring system,the function of knee joint was evaluated before and after operation.According to the condition of arounding knee joint bone fracture healing and heterotopic ossification in X-ray and CT examination and to compare the correlation of probability between heterotopic ossification and Cerus and cucumis polypeptide injection in traumatic around knee joint fracture.Results All were followed up with 12.0-24.0 months,with average(13.6±2.2)months.Activity of knee joint between group A and B were respectively(47.2±7.3)°,(46.4±6.8)°before treatment,and HSS score of group A and B were respectively(50.8±10.0)points,(51.5±9.6)points,so there were no statistically significant difference(P＞0.05).Activity of knee joint between group A and B were respectively(115.1±6.5)°,(112.1±7.8)°at the time of the last follow-up,and HSS score of group A and B were respectively(86.2±5.0)points,(85.5±7.0)points,so there were no statistically significant difference(P＞0.05).There were statistically significant difference in activity of knee joint and HSS scores in group A or in group B at the time of the last follow-up(t=13.322,12.898,10.052,10.435,P＜0.05).The incidence of heterotopic ossification was 8.99％(8/89)in group A,the incidence of heterotopic ossification was 1.64％(1/61)in group B,there were statistically significant difference between A and B(x2=10.873,P＜0.05).Conclusion During the drug treatment,there is a correlation between heterotopic ossification and Cerus and cucumis polypeptide injection used in traumatic around knee joint fracture after operation.

Objective To investigate the effect of L-carnitine injection on oxidative stress and clinical efficacy in patients with acute anterior ST segment elevated myocardial infarction ( STEMI ) and received primary percutaneous coronary intervention .Methods 70 patients with acute anterior wall STEMI undergoing primary PCI in our hospital were randomly divided into the L-carnitine treatment group and the control group.Patients in both groups received same standardized treatment including aspirin , clopidogrel, statins, ACEI/ARB etc.after PCI.Serum levels of superoxied dismutase ( SOD ) and malondialdehyde ( MDA) were measured at arrival and 7 days after PCI.Laboratory results and echocardiography finding were compared.Results (1) The serum SOD levels in both groups were elevated on day 7 after treatment (P<0.05).The serum SOD level on day 7 in the L-carnitine treatment group was higher than the control group (P<0.05).(2) The serum MDA levels in both groups were lower 7 days after treatment (P<0.05).The serum MDA level in the treatment group was lower than the control group (P<0.05).(3) Echocardiography exam on day 7 showed left ventricular end diastolic diameter ( LVEDd) and left ventricular ejection fraction (LVEF) were more significantly improved compared with the control group (P<0.05).Conclusions For AMI patients receiving primary PCI , L-carnitine injection can increase serum SOD concentration and reduce the production of MDA , showing inhibiting effects on oxidative stress and has protective effect on myocardia .

Objective To investigate the effect of tube voltage and iodine concentration of contrast medium (CM) on abdominal dynamic enhanced CT image quality.Methods Six miniature pigs underwent repeated upper abdomen dynamic contrast-enhanced CT scans in 4 scanning protocols with different concentration of CM and tube voltage,namely,protocol 1,CM with iodine concentration of 270 milligrams iodine per milliliter (mg/ml) and 80 kV tube voltage;protocol 2,270 mg/ml and 120 kV;protocol 3,370 mg/ml and 80 kV and protocol 4,370 mg/ml and 120 kV.The same iodine dose (600 mg/ml) and iodine delivery rate (IDR) (920 mg/s) were used in all protocols.The CM with iodine concentration of 270 mg/ml were injected at a flow rate of 3.4 ml/s,and 370 mg/ml CM injected at 2.5 ml/s.Image reconstruction was performed with iterative reconstruction (iDose4) in protocol 1 and 3,filtered back projection (FBP) was used in protocol 2 and 4.A subjective scoring system for image quality,image noise and sharpness was conducted by 2 radiologists independently.The measured values (peak of enhanced CT values,image noise of aorta,inferior vena cava,portal vein,hepatic vein and liver parenchyma) as well as the calculated values [their time-to-peak,signal-to-noise (SNR) and contrast-to-noise (CNR) ratios] were compared between among 4 protocols.The CT volume dose index (CDTIvol) and dose length product (DLP) were recorded from the CT console after each scanning.Factorial designed ANOVA was used for comparison of enhanced CT values of vessels and liver parenchyma,noise,SNR and CNR.The Kruskal-Wallis test was used for comparison of values among the 4 protocols,including the time-to-peak enhancement of vessels and liver parenchyma,the subjective scores of image quality indices.Result There was no significant differences in subjective scores of the image quality,image noise and image sharpness (P＞0.05).The scored were more than 3,and the images with 4 scanning protocols were all acceptable for diagnosis.There was no significant differences between protocol 1 and 3,protocol 2 and 4 in the peak enhancement CT values of aorta [(729±46) HU vs.(707±59)HU,(515±84)HU vs.(513±53)HU],inferior vena cava [(366±95)HU vs.(368±92)HU,(282±39)HU vs.(262 ± 67)HU],portal vein [(213± 18)HU vs.(201 ±29)HU,(180±21)HU vs.(176±27)HU],hepatic vein [(207±18)HU vs.(193±10)HU,(179±24)HU vs.(170±14)HU] and liver parenchyma [(128±7) HU vs.(127±4) HU,(135±5)HU vs.(135±6)HU] (P＞0.05).But the CT values of vessels (aorta,inferior vena cava,portal vein and hepatic vein) in protocol 1 and 3 were significantly higher than those in protocol 2 and 4 (P＜0.05),the CT values of liver parenchyma in protocol 1 and 3 were significantly lower than values in protocol 2 and 4 (P＜0.05).The image noises of vessels were higher in protocol 1 and 3 than noises in other protocols (P＜0.05),but there was no significant difference in liver parenchyma noise among protocols (P＞0.05).No significant differences were observed on the peak times,SNR and CNR in aorta,inferior vena cava,portal vein,hepatic vein and liver parenchyma among 4 protocols (P＞0.05).The CDTIvol and DLP were 199.67 mGy,1 597.4 mGy· cm respectively in protocol 1 and 3,585.12 mGy and 4 680.9 mGy· cm in protocol 2 and 4 (scanning with 120 kV).Conclusions CM with different iodinated concentration could achieve the same enhancement in the abdominal vessels and liver parenchyma by using the proper scan protocols,which have the same IDR and iodine dose per kilogram body weight.Higher vessel enhanced peak values were achieved when using the protocols with 80 kV tube voltage than 120 kV.By using a low dose protocol of 80 kV tube voltage with the iterative reconstruction algorithm,the quality of image can be warranted.

Objective To investigate the clinical effect on incomplete spinal injury by ganglioside intravenous injection combined with intrathecal injection.Methods From January 2011 to January 2015, seventy-nine cases with irreducible articular process interlocking of cervical spine fracture with dislocation of cervical spinal cord injury,underwent one stage anterior and posterior surgical treatment,postoperative routine use of antibiotics to prevent infection,and the hormone,dehydration to promote bone cell growth and neurotrophic drugs treatment.The patients were randomly divided into the intravenous injection group(42 cases),given intravenous injection of monalsialic acid four hexose ganglioside sodium(GM-1)40 mg/d,mecobalamin tablets 0.5 mg/time,3 times/d,30 d oral;the combined intrathecal injection group(37 cases)was given GM-1 40 mg/d,intravenous injection at 15 d after intrathecal injection,1 time a week 40 mg,with a total of 4 weeks.The degree of spinal cord injury was evaluated according to Frankel classification; cervical function was evaluated according to JOA score; bone graft fusion,stability of cervical spine and degree of spinal cord injury were evaluated by imaging.Results The operation time in the intravenous injection group and the combined intrathecal injection group were(4.15 ± 0.65)h and(4.10 ± 0.85)h,and the intraoperative blood loss was(850.50±35.10)ml and(858.60±25.20)ml,respectively,and there were no significant differences between the two groups(t=1.375,1.452,P>0.05).The total dose of GM-1 in the combined intrathecal injection group was(785.20 ± 3.28)mg,significantly higher than that in the intravenous injection group((610.55 ± 5.28) mg),the difference was statistically significant(t=12.542,P<0.05);79 patients were followed up for 12-24 months,with an average of(15.2 ± 1.3)months.The improvement rate of nerve function of the combined intrathecal injection group was(64.35±4.33)%,significantly higher than that in the intravenous injection group (55.50±5.44)%,the difference was statistically significant(t=8.813,P<0.05);the postoperative JOA scores of the intravenous injection group((13.55 ± 1.75)points)and combined intrathecal injection group((12.85 ±1.97)points)were significantly higher than those before the surgery((7.25± 0.83)points,(7.19± 0.93) points),the differences were statistically significant(P<0.05).There was no significant difference in the JOA scores between the two groups before and after the operation(P>0.05).At the last follow-up,X-ray showed bone fusion at the bone graft site,and the internal fixation was good and firm.MR showed that the degeneration signal area of the cervical spinal cord decreased in varying degrees,and edema and inflammatory reaction disappeared.Conclusion Postoperative treatment of ganglioside intravenous injection combined with intrathecal injection is safe and feasible in the treatment of incomplete cervical spinal cord injury caused by cervical fracture dislocation with irreducible articular process interlocking.

OBJECTIVETo investigate the efficacy and safety of laparoscopic fundoplication for gastroesophageal reflux disease (GERD)-related cough.

METHODSRetrospective review of 70 patients with GERD-related cough who received laparoscopic fundoplication in GERD Department of The Second Artillery General Hospital during June 2008 to June 2013 was carried out. GERD-related symptoms (reflux, heartburn, cough, expectoration, globus sensation and hoarseness) before and after surgery were compared through questionnaire, and the symptom remission rate (preoperative symptom score - postoperative symptom score)/preoperative symptom score×100%) was calculated. Complication morbidity and satisfaction degree of patients were investigated.

RESULTSThe GERD-related symptom scores of regurgitation, heartburn, cough, expectoration, globus sensation and hoarseness all significantly decreased (all P<0.01) after the anti-reflux laparoscopic fundoplication, with the corresponding symptom remission rates as (79.4±23.2)%, (82.0±21.5)%, (72.2±28.5)%, (62.6±28.9)%, (76.1±31.5)% and (70.8±39.3)% respectively. No major complication and death occurred. Five cases (7.1%) had pneumoperitoneum-related chest or neck subcutaneous emphysema, 17 cases (24.3% ) had various degree of early and late dysphagia, 6 cases (8.6%) had increased flatus and 2 cases had bloating after surgery. All the complications could be cured by appropriate treatment. Among all the patients, 16 cases (15.7%) felt very satisfied, 37 cases (52.9%) felt satisfied, 11 cases (15.7%) felt acceptable, 4 cases (5.7%) felt unsatisfied and 2 cases felt very unsatisfied with the surgery.

CONCLUSIONLaparoscopic fundoplication is safe and effective for GERD-related cough, with quite high satisfaction degree form patients.

Objective:To investigate the application value of baseline MRI multi-parametric imaging radiomics in prediction of neoadjuvant chemoradiotherapy (NCR) efficacy of rectal mucinous adenocarcinoma (RMAC).Methods:Retrospective analysis was performed in the Sixth Affiliated Hospital of Sun Yat-sen University from August 2012 to October 2018. A total of 79 patients were included in this study, including 52 males and 27 females, aged 20-78 years (median age 52 years). According to the classification criteria of pathological regression, all patients were divided into NCR responsiveness group ( n=31) and nonresponsiveness group ( n=48). And 701 imaging features of T 2WI, diffusion weighted imaging (DWI) and enhanced T 1WI images of baseline MRI were extracted, and feature subsets were selected by repeatability analysis and feature dimensionality reduction to construct the radiomics prediction model. The tumor features from baseline MRI between the NCR responsiveness group and the nonresponsiveness group were compared, and the features of P<0.05 were combined with the radiomics to construct a model. Using pathology as the gold standard, the receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficiency of the prediction model, and the area under the curve (AUC), 95% confidence interval, sensitivity and specificity were calculated, and the DeLong test was used to compare the diagnostic efficacy of different prediction models. Results:By comparing the conventional tumor imaging characteristics of the NCR responsiveness group and the nonresponsiveness group, the differences in lymph node stage and mucinous nodule status between the two groups were statistically significant (χ2 =6.040, 5.870, P<0.05). The AUC of ROC curves based on T 2WI, DWI, and enhanced T 1WI radiomics were 0.816, 0.821, and 0.819, respectively, which were higher than those of conventional tumor characteristics (lymph node staging, mucinous nodule status) (AUC=0.607), and the differences were statistically significant ( Z=-2.391, -2.580 and -2.717, P<0.05). Among the joint prediction models of T 2WI, DWI and contrast-enhanced T 1WI radiomics and conventional tumor features, the DWI combined model had the largest AUC (0.843), and there was no statistically significant difference between the three combined models (all P>0.05). Conclusion:The baseline T 2WI, DWI, and contrast-enhanced T 1WI radiomics model can be used to predict the NCR efficacy of RMAC, which is better than the predictive efficacy of conventional features, and the combination with conventional features can further improve the predictive efficacy.

Objective:To discuss the effect of exosome(Exo)microRNA-761(miR-761)on the epithelial-mesenchymal transition(EMT)process of the osteosarcoma(OS)cells by regulating tumor-associated macrophage(TAM)polarization,and to clarify its related mechanism.Methods:The miR-761 plasmid and negative control(miR-NC)plasmid were transfected into the HEK293 cells,and the non-transfected cells were regarded as control group.The transfection efficiency was detected using real-time fluorescence quantitative PCR(RT-qPCR)method.The Exo containing miR-761 was isolated,and the morphology of Exo was observed by transmission electron microscope.The concentration and size distribution of Exo samples were detected by nanoparticle analyzer,and the expression of Exo surface marker protein was detected by Western blotting method.The human monocyte leukemia THP-1 cells were stimulated with phorbol 12-myristate 13-acetate(PMA)to become the M0 macrophages,which were then treated with Exo containing miR-761 and co-cultured with the OS MG63 cells to establish the co-culture system.The experiment was divided into M0 group,TAM group,miR-761 NC group,and miR-761 Exo group.The M0 macrophages were collected from various groups,and the positive rates of M1 macrophage marker CD86 and M2 macrophage marker CD206 in various groups were detected by flow cytometry;the protein expression levels of M1 macrophage secreted factors interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)and M2 macrophage secreted factors interleukin-10(IL-10)and transforming growth factor-β1(TGF-β1)in various groups were detected by Western blotting method.The M0 macrophages were treated with Exo containing miR-761 and co-cultured with MG63 cells to establish the co-culture system.The experiment was divided into control group,TAM group,miR-NC Exo+TAM group,and miR-761 Exo+TAM group.The MG63 cells in various groups were collected,and the fluorescence intensities of E-cadherin and Vimentin in the MG63 cells in various groups were observed by immunofluorescence staining;the expression levels of E-cadherin,Vimentin,and EMT regulation-related transcription factors Twist1,Snail,and Slug proteins in the cells in various groups were detected by Western blotting method;the numbers of invasion and migration cells in various groups were detected by Transwell chamber assay.Results:The HEK293 cells containing miR-761 were successfully obtained by transfection experiments,and the Exo was isolated.Compared with M0 group,the positive rate of CD86 of the macrophages in TAM group was decreased(P＜0.05),while the positive rate of CD206 was increased(P＜0.05),the expression levels of IL-1β and TNF-α proteins were decreased(P＜0.05),while the expression levels of IL-10 and TGF-β1 proteins were increased(P＜0.05).Compared with TAM group,the positive rate of CD86 of the macrophages in miR-761 Exo group was increased(P＜0.05),while the positive rate of CD206 was decreased(P＜0.05),the expression levels of IL-1β and TNF-α proteins were increased(P＜0.05),while the expression levels of IL-10 and TGF-β1 proteins were decreased(P＜0.05).Compared with control group,the fluorescence intensity of E-cadherin in the MG63 cells in TAM group was decreased,while the fluorescence intensity of Vimentin was increased,the expression level of E-cadherin protein was decreased(P＜0.05),while the expression levels of Vimentin,Twist1,Snail,and Slug proteins were increased(P＜0.05),and the numbers of invasion and migration cells were increased(P＜0.05).Compared with TAM group,the fluorescence intensity of E-cadherin in the MG63 cells in miR-761 Exo+TAM group was increased,while the fluorescence intensity of Vimentin was decreased,the expression level of E-cadherin protein was increased(P＜0.05),while the expression levels of Vimentin,Twist1,Snail,and Slug proteins were decreased(P＜0.05),and the numbers of invasion and migration cells were decreased(P＜0.05).Conclusion:The exo-delivered miR-761 can inhibit the EMT process of the OS cells,thereby inhibiting the cell migration and cell invasion;its mechanism may be related to regulating TAM polarization.

Objective To evaluate the intraoperative identification of ectopic parathyroid tissue in the central neck region using autofluorescence point-spectral analysis(AFPSA)combined with immune colloidal gold technique(ICGT),for guiding total parathyroidectomy(TPTX)or clean parathyroidectomy(CPTX)in the management of secondary hyperparathyroidism(SHPT).Methods Retrospectively collected and compared the clinical data of 64 patients with SHPT from October 2019 to June 2023.In the observation group,TPTX was performed as the initial procedure in 36 cases,followed by sampling of suspicious targets using AFPSA in the central neck area and subsequent detection through ICGT.CPTX was then conducted if a positive result was obtained.On the other hand,the control group consisted of 28 cases where only TPTX was performed without any additional tests during surgery.The surgical data,parathyroid hormone(PTH)levels,blood calcium levels,blood phosphorus levels,alkaline phosphatase(ALP)levels,regression of clinical symptoms,changes in parathyroid function and occurrence of hypocalcemia were compared between these two groups.Results In the observation group,there were 9 cases of AFPSA-ICGT positivity,including 2 left-sided cases,4 right-sided cases,and 3 thymic cases;among these posi-tive cases,there were a total of 10 locations with mildly hyperplastic or nonhyperplastic microscopic parathyroid tissue.The difference in the number of total parathyroid glands removed(including ectopic)between the two groups was statistically significant(P<0.05).At both 3 and 6 months postoperatively,ALP levels in the observation group were significantly lower than those in the control group(P<0.01 and P<0.001 respectively);at 6 months postoperatively,differences in PTH and blood phosphorus levels between the two groups were also statistically significant(P<0.05 and P<0.001 respectively).Joint bone pain and skin itching recurred in some patients within the control group at six months after surgery(P<0.05),whereas recurrence of SHPT was less frequent within the observation group compared to controls(P<0.05);however,no statistically significant differences were observed regarding postoperative hypoparathyroidism or hyperparathyroidism as well as hypocalcemia between either groups.Conclusion The AFPSA-ICGT intraoperative test can be utilized to guide surgery for SHPT,enabling accurate and efficient identification as well as safe targeting of parathyroid tissues that may not exhibit obvious hyperplasia in the central cervical region.

ObjectiveTo observe the clinical effect of modified Jichuanjian on senile patients with slow transit constipation of spleen-kidney yang deficiency syndrome and the influence on brain-gut peptide. MethodA total of 150 senile patients with slow transit constipation were randomized into control group （75 cases） and observation group （75 cases） with the random number table method. The observation group was given modified Jichuanjian （oral， 1 dose/day， 4 weeks）， and the control group was treated with Biantong Capsules （oral， 3 capsules/time， twice/day， 4 weeks）. Data before and after treatment were recorded， including the score of major constipation symptoms， score of Patient Assessment of Constipation Quality of Life （PAC-QOL）， TCM syndrome score， spontaneous complete bowel movements （SCBM）， colonic transit test， serum 5-hydroxytryptamine （5-HT）， 5-HT 4 receptor （5-HT₄R）， somatostatin （SS）， and vasoactive intestinal peptide （VIP）， and recurrence. ResultThe total effective rate of the observation group was 93.06% （67/72）， as compared with the 74.65% （53/71） in the control group （χ²=8.974 6， P<0.01）. After treatment， the scores of major constipation symptoms， scores of four dimensions of PAC-QOL， total score of PAC-QOL， and TCM syndrome score were lower than those before treatment in the two groups （P<0.01）， and lower in the observation group than in the control group （P<0.01）. The SCBM in the observation group were more than those in the control group at the 2^nd， 3^rd， 4^th weeks after treatment （P<0.01）. The proportions of residual markers at 24， 48， 72 h after treatment were smaller than those before treatment in the two groups （P<0.01）， and smaller in the observation group than in the control group （P<0.01）. After treatment， the levels of serum 5-HT and 5-HT₄R were higher （P<0.01） and the levels of serum SS and VIP were lower （P<0.01） than those before treatment in the two groups. In addition， the levels of serum 5-HT and 5-HT₄R in the observation group were higher （P<0.01） and the levels of serum SS and VIP were lower （P<0.01） in the observation group than in the control group. The recurrence in the observation group was 29.85% （20/67） in comparison with the 58.49% （31/53） in the control group （χ²=9.932 4， P<0.01）. ConclusionModified Jichuanjian is effective for senile patients with slow transit constipation of spleen-kidney yang deficiency syndrome， which can alleviate clinical symptoms， improve quality of life， regulate the level of serum brain-gut peptide， improve the colonic transit function， increase SCBM， and reduce the recurrence.